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Altered expression of circular RNA in human dental pulp cells during odontogenic differentiation

机译:牙源性分化过程中人牙髓细胞中环状RNA表达的改变

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摘要

The alterations in expression and function of circular RNA (circRNA) in human dental pulp cells (hDPCs) during odontogenic differentiation were investigated. To induce odontogenic differentiation, hDPCs (passage 3) were cultured for 14 days in odontogenic induction medium. circRNA high-throughput sequencing was performed using Illumina HiSeqseq™ 2000. Subsequently, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes analyses were used to evaluate the bio-functions of the identified circRNAs. To validate the results of circRNA sequencing, reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed for two selected differentially expressed circRNAs. The RNA sequencing results revealed that 1,314 and 1,780 circRNAs were upregulated and downregulated, respectively, during odontogenic induction. Their predicted target miRNAs and genes are involved in several biological functions and signaling pathways, including the mitogen-associated protein kinase signaling pathway. The RT-qPCR results of the two selected circRNAs (hsa_circ_0015260 and hsa_circ_0006984) were consistent with the expression trend obtained using high-throughput sequencing. The results of the present study add to the current understanding of the regulatory mechanisms underlying hDPCs differentiation.
机译:研究了在牙源性分化过程中人牙髓细胞(hDPC)中环状RNA(circRNA)的表达和功能的变化。为了诱导牙源性分化,将hDPC(第3代)在牙源性诱导培养基中培养14天。使用Illumina HiSeqseq™2000进行circRNA高通量测序。随后,使用基因本体论和《京都议定书》的基因与基因组百科全书分析评估鉴定出的circRNA的生物功能。为了验证circRNA测序的结果,对两个选定的差异表达circRNA进行了逆转录定量聚合酶链反应(RT-qPCR)。 RNA测序结果表明,在牙源性诱导过程中,分别有1314和1780个circRNA上调和下调。他们预测的靶标miRNA和基因参与了几种生物学功能和信号传导途径,包括有丝分裂原相关的蛋白激酶信号传导途径。两种选择的circRNA(hsa_circ_0015260和hsa_circ_0006984)的RT-qPCR结果与使用高通量测序获得的表达趋势一致。本研究的结果增加了目前对hDPCs分化的调控机制的了解。

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